Sperm acquires capacity of motility and fertility during the process of semen coagulation and liquefaction. The main coagulative protein is Semenogelin I (Sg I), specifically produced by seminal vesicles, and then decomposed by prostate specific antigens (PSA) in sperm liquefaction into a series of small fragments. These fragments, with a variety of physiological functions, are very important for the regulation of sperm capacity acquisition and progressive movement.
Humans ; Male ; Seminal Vesicle Secretory Proteins ; physiology ; Seminal Vesicles ; metabolism ; physiology

Semen liquefaction and sperm capacitation are the key processes for sperm to acquire forward movement ability. In these processes, semenogelin plays a vital role by directly participating in the formation of semen coagulation, collaborating with other protease and metal ions from the male reproductive tract, and then reacting with the surface of sperm cells, finally involved in the regulation of these processes and ensuring sperm's acquisition of forward movement ability.
Humans ; Male ; Semen ; chemistry ; Seminal Vesicle Secretory Proteins ; physiology ; Sperm Motility

OBJECTIVETo evaluate the correlation of epididymal protease inhibitor(Eppin) and Semenogelin(Sg) on human ejaculated spermatozoa.

METHODSThe experimental approaches include: (1) Immunoprecipitation of Eppin with anti-Eppin from semen; (2) Colocalization of Eppin and Sg by immunofluorescence; (3) Immunoprecipitation of rEppin and rSg;(4) Far-Western blotting of rEppin and rSg;(5) Competition of saturated 125I-rSg binding to rEppin with unlabeled Sg, and direct binding of 125I-rSg to rEppin on a blot; (6) Autoradiography of 125I-rSg with rEppin.

RESULTSEppin-Sg complex present on the surface of human ejaculated spermatozoa, Cys-239 is the only cystein for rEppin binding rSg. Reduction and carboxymethylation of Cys-239 blocks binding of 125I-rEppin to rSg.

CONCLUSIONOur study demonstrates that Eppin and Sg bind to each other on human ejaculated spermatozoa. A disulfide linkage occurs between Sg and Eppin, indicating the specificity of binding.

Humans ; Male ; Protein Binding ; Proteinase Inhibitory Proteins, Secretory ; Proteins ; chemistry ; metabolism ; Recombinant Proteins ; Seminal Vesicle Secretory Proteins ; chemistry ; metabolism ; Spermatozoa ; metabolism

Semenogelin I (Sg I) and the fragments of peptides hydrolyzed from Sg I by prostate-specific antigen have multiple biological activities. There exists a controversy over the inhibitory effect of the key fragment on sperm motility. This article focuses on the sperm-inhibiting and antibacterial activities of the fragments of Sg I-derived peptides and illustrates the supposition concerning the most controversial aspect. A deeper insight into the action mechanisms of Sg I-derived peptides may help improve the methods of sperm screening and provide a new perspective in the management of asthenozoospermia and urinary tract infection.
Anti-Bacterial Agents ; Humans ; Male ; Semen ; drug effects ; Seminal Vesicle Secretory Proteins ; genetics ; physiology ; Spermatozoa ; drug effects

The epididymal protease inhibitor (Eppin) abounds in human semen and on the surface of human spermatozoa, specifically produced by the testis and epididymis. Recombinant Eppin has effected infertility in the immunized monkey and promises to be an effective vaccine for human immunocontraception. This article reviews the advances in the studies of Eppin gene and protein construction and its molecular mechanism of causing immunologic infertility and regulating PSA hydrolysis of Semenogelin.
Animals ; Humans ; Infertility, Male ; immunology ; Male ; Mice ; Primates ; Proteinase Inhibitory Proteins, Secretory ; chemistry ; genetics ; immunology ; physiology ; Seminal Vesicle Secretory Proteins ; physiology ; Vaccines, Contraceptive

Phosphoinositide-specific Phospholipase C (PI-PLC) is a second messenger of signal transducer on cell membrane. In the previous study, PLC of bovine brain has been purified three isozymes. In this paper, uterus and seminal vesicle have been purified. Two peaks of PI-PLC activity were resolved when bovine uterus and seminal vesicle proteins were chromatographed on a DEAE and phenyl TSK 5PW HPLC column. Each two peak was compared with PI-PLC I, II and III from bovine brain and we got the retention time on HPLC. The peak fractions with PLC activity were tested homogeneity with brain PLC monoclonal antibodies (Mab). Mab-labeled affigels were bounded in the range of 73.8%~97.5% with PLC I, II and III. Homogeneity of fractions were revealed that DEAE F-1 and phenyl F-1-I were highest level of PLC III in uterus and seminal vesicle and DEAE F-2 and phenyl F-2-I were mixed PLC I and II.
Antibodies, Monoclonal ; Brain* ; Cell Membrane ; Chromatography, High Pressure Liquid ; Isoenzymes* ; Phospholipases* ; Second Messenger Systems ; Seminal Vesicle Secretory Proteins ; Seminal Vesicles* ; Transducers ; Type C Phospholipases* ; Uterus*

Sperm membrane proteins play a key role in spermatogenesis, sperm maturation and sperm-oocyte interaction. A deeper research would shed new light on the molecular mechanisms of spermatogenesis, sperm maturation and fertilization. In recent years, with the extensive application of a variety of current molecular biological methods and bioinformatics to reproductive medicine, some sperm membrane proteins found previously have been cloned and sequenced. Furthermore, new sperm membrane proteins, being found continuously, will make a solid foundation for the development of contraceptive vaccine as well as for the investigation into the mechanism of fertilization at levels of gene and protein. This article reviews the current progress in the researches on sperm membrane proteins.
Adaptor Proteins, Signal Transducing ; physiology ; Animals ; Humans ; Male ; Membrane Proteins ; physiology ; Seminal Vesicle Secretory Proteins ; physiology ; Spermatogenesis ; physiology ; Spermatozoa ; cytology ; growth & development

OBJECTIVETo study the inhibition activity of Semenogelin (Sg) and its different peptides to human spermatozoa.

METHODSHuman Sg DNA and its N-terminal Sg and C-terminal Sg DNA were cloned into PET-100 vector. Positive colonies were screened and transformed into E. Coli BL21 (DE3). Recombinant Sg and its peptides were induced and expressed in high competent E. coli BL21 (DE3) , and purified by 50% Ni-NTA column. Inhibition activity assay was done by adding 4 different concentrations of semenogelin and its two peptides, 0, 1, 5 and 10 ng/microl, to human spermatozoa.

RESULTSThe peptide of Semenogelin that inhibits the activity of human spermatozoa was located in its N-terminal fragment. C-terminal Sg did not inhibit the activity of spermatozoa.

CONCLUSIONN-terminal Sg is the inhibition peptide of the whole molecular Sg. During semen liquefaction, this peptide should be cut off from the surface of human spermatozoa before they move forward.

Cloning, Molecular ; Dose-Response Relationship, Drug ; Escherichia coli ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; pharmacology ; Seminal Vesicle Secretory Proteins ; biosynthesis ; genetics ; pharmacology ; Sperm Motility ; drug effects

During liquefaction of the ejaculate, the semen coagulum proteins semenogelin I (SEMG1) and semenogelin II (SEMG2) are degraded to low molecular mass fragments by kallikrein-related peptidase 3 (KLK3), also known as prostate-specific antigen. Semenogelin molecules initiate their own destruction by chelating Zn(2+) that normally would completely inhibit the proteolytic activity of KLK3. In a similar way, semenogelins might regulate the activity of kallikrein-related peptidases in the epididymis, something that might be of importance for the maturation of spermatozoa or generation of anti-bacterial peptides. Studies on the evolution of semen coagulum proteins have revealed that most of them carry an exon that displays a rapid and unusual evolution. As a consequence, homologous proteins in rodents and primates show almost no conservation in primary structure. Further studies on their evolution suggest that the progenitor of the semen coagulum proteins probably was a protease inhibitor that might have displayed antimicrobial activity. The semenogelin locus on chromosome 20 contains at least 17 homologous genes encoding probable protease inhibitors with homology to semen coagulum proteins. All of these are highly expressed in the epididymis where they, similar to the semenogelins, could affect the maturation of spermatozoa or display antibacterial properties.
Animals ; Centromere ; Chromosome Mapping ; Chromosomes, Human, Pair 20 ; Ejaculation ; Epididymis ; physiology ; Evolution, Molecular ; Gene Expression Regulation ; Humans ; Male ; Primates ; Semen ; physiology ; Seminal Vesicle Secretory Proteins ; genetics

AIMTo study the molecular mechanism of epididymal protease inhibitor (Eppin) modulating the process of prostate specific antigen (PSA) digesting semenogelin (Sg).

METHODSHuman Sg cDNA (nucleotides 82-849) and Eppin cDNA (nucleotides 70-723) were generated by polymerase chain reaction (PCR) and cloned into pET-100D/TOPO. Recombinant Eppin and Sg (rEppin and rSg) were produced by BL21 (DE3). The association of Eppin with Sg was studied by far-western immunoblot and radioautography. In vitro the digestion of rSg by PSA in the presence or absence of rEppin was studied. The effect of anti-Q20E (N-terminal) and C-terminal of Eppin on Eppin-Sg binding was monitored.

RESULTSEppin binds Sg on the surface of human spermatozoa with the C-terminal of Eppin (amino acids 75-133). rSg was digested with PSA and many low molecular weight fragments were produced. When rEppin is bound to rSg, then digested by PSA, incomplete digestion and a 15-kDa fragment results. Antibody binding to the N-terminal of rEppin did not affect rSg digestion. Addition of antibodies to the C-terminal of rEppin inhibited the modulating effect of rEppin.

CONCLUSIONEppin protects a 15-kDa fragment of rSg from hydrolysis by PSA.

Animals ; Antibodies ; pharmacology ; Autoradiography ; Humans ; Hydrolysis ; Male ; Prostate-Specific Antigen ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; genetics ; immunology ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Semen ; cytology ; metabolism ; Seminal Vesicle Secretory Proteins ; metabolism ; Spermatozoa ; metabolism